J. Biochem, 1986, Vol. 99, No. 5 1465-1472
© 1986 Japanese Biochemical Society
research-article |
The Initial Phosphate Burst in ATP Hydrolysis by Myosin and Subfragment-1 as Studied by a Modified Malachite Green Method for Determination of Inorganic Phosphate
*Department of Biochemistry Okayama, Okayama 700
**Department of Microbiology, Okayama University Dental School Okayama, Okayama 700
***Department of Physiology, Medical College of Oita Oita 879–56
1 To whom all inquiries should be addressed
The Malachite Green method for determination of inorganic phosphate (P1 (Itaya K. & Ui, M. (1966) Clin. Chim. Acta 14, 361–366) was modified to measure P in the range of 0.2–15 nmol per ml of ATPase reaction mixture. An ATPase reaction mixture is quenched with an equal volume of 0.6 M PCA; the supernatant after centrifugation is mixed with an equal volume of the Malachite Green/molybdate reagent containing 2 g of sodium molybdate, 0.3 g of Malachite Green and 0.5 g of Triton X-100 or Sterox SE in 1 liter of 0.7 M HCl, and the absorbance at 650 nm is then measured after a 35–40 mm incubation at 25°C.
Owing to the high sensitivity and simplicity of the modified method, the slow time course of myosin ATP hydrolysis in the presence of Mg2+ and the size of initial phosphate burst can be determined accurately using relatively low concentrations of native myosin and its subfragment-1. The phosphate burst size varied with changes in pH, ionic strength, and temperature. A typical value was 0.8–0.9 mol per site in 0.1 M KCl, 10 mr MgCl2 pH 8.0 at 25°C for fresh enzyme preparations.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  N. Albet-Torres, M. J. Bloemink, T. Barman, R. Candau, K. Frolander, M. A. Geeves, K. Golker, C. Herrmann, C. Lionne, C. Piperio, et al. Drug Effect Unveils Inter-head Cooperativity and Strain-dependent ADP Release in Fast Skeletal Actomyosin J. Biol. Chem., August 21, 2009; 284(34): 22926 - 22937. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Kobayashi, S. E. Patrick, and M. Kobayashi Ala Scanning of the Inhibitory Region of Cardiac Troponin I J. Biol. Chem., July 24, 2009; 284(30): 20052 - 20060. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. H. Wang, H. Tanaka, X. Qin, T. Zhao, L.-H. Ye, T. Okagaki, T. Katayama, A. Nakamura, R. Ishikawa, S. E. Thatcher, et al. Blebbistatin inhibits the chemotaxis of vascular smooth muscle cells by disrupting the myosin II-actin interaction Am J Physiol Heart Circ Physiol, May 1, 2008; 294(5): H2060 - H2068. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. Funabara, C. Hamamoto, K. Yamamoto, A. Inoue, M. Ueda, R. Osawa, S. Kanoh, D. J. Hartshorne, S. Suzuki, and S. Watabe Unphosphorylated twitchin forms a complex with actin and myosin that may contribute to tension maintenance in catch J. Exp. Biol., December 15, 2007; 210(24): 4399 - 4410. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Sakata, Y. Watanabe, J. Usukura, and I. Mabuchi Characterization of Native Myosin VI Isolated from Sea Urchin Eggs J. Biochem., October 1, 2007; 142(4): 481 - 490. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Takaine and I. Mabuchi Properties of Actin from the Fission Yeast Schizosaccharomyces pombe and Interaction with Fission Yeast Profilin J. Biol. Chem., July 27, 2007; 282(30): 21683 - 21694. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. Hachikubo, K. Ito, J. Schiefelbein, D. J. Manstein, and K. Yamamoto Enzymatic Activity and Motility of Recombinant Arabidopsis Myosin XI, MYA1 Plant Cell Physiol., June 1, 2007; 48(6): 886 - 891. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C.-S. Liang, A. Kobiyama, A. Shimizu, T. Sasaki, S. Asakawa, N. Shimizu, and S. Watabe Fast skeletal muscle myosin heavy chain gene cluster of medaka Oryzias latipes enrolled in temperature adaptation Physiol Genomics, April 24, 2007; 29(2): 201 - 214. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Iwai, D. Hanamoto, and S. Chaen A Point Mutation in the SH1 Helix Alters Elasticity and Thermal Stability of Myosin II J. Biol. Chem., October 13, 2006; 281(41): 30736 - 30744. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Kobayashi and R. J. Solaro Increased Ca2+ Affinity of Cardiac Thin Filaments Reconstituted with Cardiomyopathy-related Mutant Cardiac Troponin I J. Biol. Chem., May 12, 2006; 281(19): 13471 - 13477. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Onishi, S. V. Mikhailenko, and M. F. Morales Toward understanding actin activation of myosin ATPase: The role of myosin surface loops PNAS, April 18, 2006; 103(16): 6136 - 6141. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Hook, A. Mikami, B. Shafer, B. T. Chait, S. S. Rosenfeld, and R. B. Vallee Long Range Allosteric Control of Cytoplasmic Dynein ATPase Activity by the Stalk and C-terminal Domains J. Biol. Chem., September 23, 2005; 280(38): 33045 - 33054. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Morii, T. Shimizu, N. Mizuno, M. Edamatsu, K. Ogawa, Y. Shimizu, and Y. Y. Toyoshima Removal of Tightly Bound ADP Induces Distinct Structural Changes of the Two Tryptophan-Containing Regions of the ncd Motor Domain J. Biochem., July 1, 2005; 138(1): 95 - 104. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Zhong, D. H. Burk, C. J. Nairn, A. Wood-Jones, W. H. Morrison III, and Z.-H. Ye Mutation of SAC1, an Arabidopsis SAC Domain Phosphoinositide Phosphatase, Causes Alterations in Cell Morphogenesis, Cell Wall Synthesis, and Actin Organization PLANT CELL, May 1, 2005; 17(5): 1449 - 1466. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Burton, H. White, and J. Sleep Kinetics of muscle contraction and actomyosin NTP hydrolysis from rabbit using a series of metal-nucleotide substrates J. Physiol., March 15, 2005; 563(3): 689 - 711. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Muto, H. Sakai, and K. Kaseda Long-range cooperative binding of kinesin to a microtubule in the presence of ATP J. Cell Biol., February 28, 2005; 168(5): 691 - 696. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Zhong, D. H. Burk, W. H. Morrison III, and Z.-H. Ye FRAGILE FIBER3, an Arabidopsis Gene Encoding a Type II Inositol Polyphosphate 5-Phosphatase, Is Required for Secondary Wall Synthesis and Actin Organization in Fiber Cells PLANT CELL, December 1, 2004; 16(12): 3242 - 3259. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Zhong and Z.-H. Ye Molecular and Biochemical Characterization of Three WD-Repeat-Domain-containing Inositol Polyphosphate 5-Phosphatases in Arabidopsis thaliana Plant Cell Physiol., November 15, 2004; 45(11): 1720 - 1728. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Ajtai, S. P. Garamszegi, S. Watanabe, M. Ikebe, and T. P. Burghardt The Myosin Cardiac Loop Participates Functionally in the Actomyosin Interaction J. Biol. Chem., May 28, 2004; 279(22): 23415 - 23421. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Al-Bassam, Y. Cui, D. Klopfenstein, B. O. Carragher, R. D. Vale, and R. A. Milligan Distinct conformations of the kinesin Unc104 neck regulate a monomer to dimer motor transition J. Cell Biol., November 24, 2003; 163(4): 743 - 753. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Ito, T. Q. P. Uyeda, Y. Suzuki, K. Sutoh, and K. Yamamoto Requirement of Domain-Domain Interaction for Conformational Change and Functional ATP Hydrolysis in Myosin J. Biol. Chem., August 15, 2003; 278(33): 31049 - 31057. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Hachikubo, K. Ito, and K. Yamamoto Roles of the Hydrophobic Triplet in the Motor Domain of Myosin in the Interaction between Myosin and Actin J. Biochem., July 1, 2003; 134(1): 165 - 171. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Kaseda, H. Higuchi, and K. Hirose From the Cover: Coordination of kinesin's two heads studied with mutant heterodimers PNAS, December 10, 2002; 99(25): 16058 - 16063. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Onishi, T. Ohki, N. Mochizuki, and M. F. Morales Early stages of energy transduction by myosin: Roles of Arg in Switch I, of Glu in Switch II, and of the salt-bridge between them PNAS, November 26, 2002; 99(24): 15339 - 15344. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 I. P. Udovichenko, D. Gibbs, and D. S. Williams Actin-based motor properties of native myosin VIIa J. Cell Sci., January 15, 2002; 115(2): 445 - 450. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Buster, K. McNally, and F. J. McNally Katanin inhibition prevents the redistribution of {gamma}-tubulin at mitosis J. Cell Sci., January 3, 2002; 115(5): 1083 - 1092. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. L. Wilson, M. S. Ou, H. C. Aldrich, and J. Maupin-Furlow Biochemical and Physical Properties of the Methanococcus jannaschii 20S Proteasome and PAN, a Homolog of the ATPase (Rpt) Subunits of the Eucaryal 26S Proteasome J. Bacteriol., March 15, 2000; 182(6): 1680 - 1692. [Abstract] [Full Text]
|
|
|
|
|
|
J. H. Gerson, E. Bobkova, E. Homsher, and E. Reisler Role of Residues 311/312 in Actin-Tropomyosin Interaction. IN VITRO MOTILITY STUDY USING YEAST ACTIN MUTANT E311A/R312A J. Biol. Chem., June 18, 1999; 274(25): 17545 - 17550. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L.-H. Ye, H. Kishi, A. Nakamura, T. Okagaki, T. Tanaka, K. Oiwa, and K. Kohama Myosin light-chain kinase of smooth muscle stimulates myosin ATPase activity without phosphorylating myosin light chain PNAS, June 8, 1999; 96(12): 6666 - 6671. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. M. Apfel, B. Takács, M. Fountoulakis, M. Stieger, and W. Keck Use of Genomics To Identify Bacterial Undecaprenyl Pyrophosphate Synthetase: Cloning, Expression, and Characterization of the Essential uppS Gene J. Bacteriol., January 15, 1999; 181(2): 483 - 492. [Abstract] [Full Text]
|
|
|
|
|
|
X. Liu, K. Ito, S. Morimoto, A. Hikkoshi-Iwane, T. Yanagida, and T. Q. P. Uyeda Filament structure as an essential factor for regulation of Dictyostelium myosin by regulatory light chain phosphorylation PNAS, November 24, 1998; 95(24): 14124 - 14129. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Sasaki, T. Shimada, and K. Sutoh Mutational Analysis of the Switch II Loop of Dictyostelium Myosin II J. Biol. Chem., August 7, 1998; 273(32): 20334 - 20340. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L.-H. Ye, K. Hayakawa, H. Kishi, M. Imamura, A. Nakamura, T. Okagaki, T. Takagi, A. Iwata, T. Tanaka, and K. Kohama The Structure and Function of the Actin-binding Domain of Myosin Light Chain Kinase of Smooth Muscle J. Biol. Chem., December 19, 1997; 272(51): 32182 - 32189. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Yumura and T. Q.P. Uyeda Transport of Myosin II to the Equatorial Region without Its Own Motor Activity in Mitotic Dictyostelium Cells Mol. Biol. Cell, October 1, 1997; 8(10): 2089 - 2099. [Abstract] [Full Text]
|
|
|
|
|
|
L. Feng, E. Kim, W.-L. Lee, C. J. Miller, B. Kuang, E. Reisler, and P. A. Rubenstein Fluorescence Probing of Yeast Actin Subdomain 3/4 Hydrophobic Loop 262-274. ACTIN-ACTIN AND ACTIN-MYOSIN INTERACTIONS IN ACTIN FILAMENTS J. Biol. Chem., July 4, 1997; 272(27): 16829 - 16837. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Tucker and L. S.B. Goldstein Probing the Kinesin-Microtubule Interaction J. Biol. Chem., April 4, 1997; 272(14): 9481 - 9488. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Hayashi, R. Ishikawa, L.-H. Ye, X.-L. He, K. Takata, K. Kohama, and T. Shirao Modulatory Role of Drebrin on the Cytoskeleton within Dendritic Spines in the Rat Cerebral Cortex J. Neurosci., November 15, 1996; 16(22): 7161 - 7170. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Kamimoto, T. Zama, R. Aoki, Y. Muro, and M. Hagiwara Identification of a Novel Kinesin-related Protein, KRMP1, as a Target for Mitotic Peptidyl-prolyl Isomerase Pin1 J. Biol. Chem., September 28, 2001; 276(40): 37520 - 37528. [Abstract] [Full Text] [PDF]
|
|